By combining biochemical studies (including protein-protein and protein-lipid binding studies of native or of purified, recombinant components) with fluorescence-based microscopy techniques applied to cellular in vivo systems we were able to elaborate novel and important aspects about how cargo recognition, coat polymerization and membrane deformation are coordinated during the generation of transport carriers.

The formation of transport carriers on different subcellular membranes is coupled to the local generation of distinct species of phosphoinositides (PIs). PI(4,5)P2 (cyan) supports endocytic events at the plasma membrane, whereas PI(4)P (green) and PI(3)P (red) are predominantly synthesized during budding from Golgi membranes and from organelles of endosomal origin, respectively. Different PIs in turn facilitate membrane translocation of adaptor and accessory proteins, which are crucial for cargo recognition, membrane deformation and budding.